A definitive role of Shp-2 tyrosine phosphatase in mediating embryonic stem cell differentiation and hematopoiesis

Rebecca Chan, Scott A. Johnson, Yanjun Li, Mervin Yoder, Gen Sheng Feng

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Homozygous mutant (Shp-2Δ46-110) embryonic stem (ES) cells exhibit decreased hematopoiesis; however, the point at which Shp-2 is critical for ES cell differentiation to hematopoietic cells is unknown. We characterized the differentiation defect of Shp-2Δ46-110 ES cells by examining early points of differentiation, conducting leukemia inhibitory factor (LIF)-stimulated biochemical analysis, and performing in vitro reconstitution studies with wild-type (WT) Shp-2. ES cell in vitro differentiation assays were used to compare the differentiation of WT, Shp-2Δ46-110, and reconstituted ES cells to mesoderm, by measuring brachyury expression, to hemangioblasts, by measuring blast colony-forming cell (BL-CFC) formation and flk-1 expression, and to hematopoietic progenitor colony-forming cells, by performing secondary plating assays. LIF-stimulated phospho-Stat3 (known to be critical for ES cell self-renewal and maintenance of an undifferentiated state) and phospho-Erk levels were examined by immunoblotting. ES cell survival, using annexin V staining, and secondary embryoid body (EB) formation were also evaluated. Differentiation to both mesoderm and hemangioblasts was lower in Shp-2 Δ46-110 cells compared to WT cells. On reconstitution with WT Shp-2, expression of brachyury and flk-1 and differentiation to hemangioblasts and primitive and definitive hematopoietic progenitors were restored. LIF-stimulated phospho-Stat3 levels were higher, whereas phospho-Erk levels were lower in Shp-2Δ46-110 ES cells than in WT and reconstituted cells. The increased phospho-Stat3 levels correlated with increased Shp-2Δ46-110 ES cell secondary EB formation and survival. We conclude that normal Shp-2 function is critical for the initial step of ES cell differentiation to mesoderm and to hemangioblasts and acts within the LIF-gp130-Stat3 pathway to maintain a proper balance of ES cell differentiation, pluripotency, and apoptosis.

Original languageEnglish (US)
Pages (from-to)2074-2080
Number of pages7
JournalBlood
Volume102
Issue number6
DOIs
StatePublished - Sep 15 2003
Externally publishedYes

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Non-Receptor Type 11 Protein Tyrosine Phosphatase
Hematopoiesis
Embryonic Stem Cells
Stem cells
Cell Differentiation
Hemangioblasts
Leukemia Inhibitory Factor
Mesoderm
Embryoid Bodies
Assays
Annexin A5
Plating
Immunoblotting

ASJC Scopus subject areas

  • Hematology

Cite this

A definitive role of Shp-2 tyrosine phosphatase in mediating embryonic stem cell differentiation and hematopoiesis. / Chan, Rebecca; Johnson, Scott A.; Li, Yanjun; Yoder, Mervin; Feng, Gen Sheng.

In: Blood, Vol. 102, No. 6, 15.09.2003, p. 2074-2080.

Research output: Contribution to journalArticle

Chan, Rebecca ; Johnson, Scott A. ; Li, Yanjun ; Yoder, Mervin ; Feng, Gen Sheng. / A definitive role of Shp-2 tyrosine phosphatase in mediating embryonic stem cell differentiation and hematopoiesis. In: Blood. 2003 ; Vol. 102, No. 6. pp. 2074-2080.
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